Registration Dossier

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study performed under GLP. All relevant validity criteria were met.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
inspected June 2015; signature: September 2015

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Physical state: Liquid.
- Storage condition of test material: Room temperature in the dark for approximately 24 hours after receipt, thereafter approximately 4 °C in the dark
- Other: Colourless liquid

Test animals / tissue source

Species:
other: bovine
Strain:
not specified

Test system

Vehicle:
unchanged (no vehicle)
Controls:
other: negative (0.9% w/v sodium chloride solution) and positive (ethanol; > 99.8% purity) controls were used in this study.
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.75 mL
- Concentration (if solution): undiluted
Duration of treatment / exposure:
10 minutes at 32 ± 1ºC.
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): At the end of the exposure period the test item and control items were removed from the anterior chamber and the cornea was rinsed three times with fresh complete EMEM containing phenol red before a final rinse with complete EMEM without phenol red. The anterior chamber was refilled with fresh complete EMEM without phenol red. A post-treatment opacity reading was taken and each cornea was visually observed. The holders were incubated, anterior chamber facing forward, at 32 ± 1 ºC for 120 minutes. After incubation the holders were removed from the incubator, the medium from both chambers was replaced with fresh complete EMEM and a final opacity reading was taken. Each cornea was visually observed.
- Time after start of exposure: 10 minutes

SCORING SYSTEM: The mean opacity and mean permeability values (OD492) were used for each treatment group to calculate an in vitro score:
In vitro irritancy score (IVIS) = mean opacity value + (15 x mean OD492 value). A test item that induces an In Vitro Irritancy Score >/=55.1 is defined as an ocular corrosive or severe irritant. A test item with an IVIS

Results and discussion

In vivo

Results
Irritation parameter:
other: In vitro irritancy score
Basis:
mean
Score:
0.8
Reversibility:
not specified
Irritant / corrosive response data:
The corneas treated with the test item were clear post treatment and post incubation. The corneas treated with the negative control item were clear post treatment and post incubation. The corneas treated with the positive control item were cloudy post treatment and post incubation.

Any other information on results incl. tables

Table 1. Individual and Mean Corneal Opacity and Permeability Measurements

Treatment

Cornea Number

Opacity

Permeability (OD)

In Vitro Irritancy Score

Pre-Treatment

Post-Treatment

Post Incubation

Post-Incubation - Pre‑Treatment

Corrected Value

 

Corrected Value

Negative Control

3

4

6

6

2

-

0.036

-

-

7

4

4

4

0

-

0.111

-

-

9

4

5

4

0

-

0.048

-

-

-

-

-

-

0.7 #1

-

0.065 #2

-

1.6

Positive Control

1

3

35

36

33

32.3

0.945

0.880

-

2

5

36

37

32

31.3

0.967

0.902

-

4

5

30

28

23

22.3

0.690

0.625

-

-

-

-

-

-

28.7 #3

-

0.802 #3

40.7

Test Item

5

5

3

3

-2

0.0

0.074

0.009

-

6

5

6

8

3

2.3

0.037

0.000

-

8

3

4

3

0

0.0

0.027

0.000

-

-

-

-

-

-

0.8 #3

-

0.003 #3

0.8

OD = Optical Density

#1 = Mean of post-incubation

#2 = Mean permeability

#3 = Mean corrected value

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of this study the test substance is not considered to be irritant in the in vitro eye corrosion/irritation test using Bovine Corneal Opacity and Permeability model. In vitro irritancy score (IVIS) was < 3.0 in the prediction model.
Executive summary:

The study was performed according to OECD TG 437 and EU Method B.47 to assess the eye irritancy potential in accordance with GLP of the test material in isolated bovine corneas. The purpose of this test was to identify test items that can induce serious eye damage and to identify test items not requiring classification for eye irritation or serious eye damage. The Bovine Corneal Opacity and Permeability (BCOP) test method is an organotypic model that provides short term maintenance of normal physiological and biochemical function of the bovine cornea in vitro. In this test method, damage by the test item is assessed by quantitative measurements of changes in corneal opacity and permeability. The ocular irritancy of the test substance was tested through topical application for 10 ± 1 minutes. The negative control responses for opacity and permeability were less than the upper limits of the laboratory historical range indicating that the negative control did not induce irritancy on the corneas. The mean in vitro irritancy score of the positive control (ethanol), was 40.7 and was within the historical positive control data range (29.6 to 52.09). The test substance did not induce ocular irritation through both endpoints, resulting in a mean in vitro irritancy score (IVIS) of 0.8 after 10 minutes of treatment. Since the IVIS was < 3.0 the test substance was predicated as not irritating to the eye. Under the conditions of this study the test material is not considered to be a irritant or corrosive in the Bovine Corneal Opacity and Permeability test.